Development and validation of reverse-transcription cross-priming amplification-based lateral flow assay for the detection of infectious hematopoietic necrosis virus.

Autor: Choi HD; Department of Aquatic Life Medicine, Pukyong National University, Busan, Republic of Korea., Baek EJ; Department of Aquatic Life Medicine, Pukyong National University, Busan, Republic of Korea., Hong S; Department of Aquatic Life Medicine, Gangneung-Wonju National University, Gangneung, Republic of Korea., Kim YC; Department of Aquatic Life Medicine, Gangneung-Wonju National University, Gangneung, Republic of Korea., Jeong JM; Aquatic Disease Control Division, National Fishery Products Quality Management Service (NFQS), Republic of Korea., Kwon MG; Aquatic Disease Control Division, National Fishery Products Quality Management Service (NFQS), Republic of Korea., Il Kim K; Department of Aquatic Life Medicine, Pukyong National University, Busan, Republic of Korea. Electronic address: kimki@pknu.ac.kr.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2024 Sep; Vol. 329, pp. 115008. Date of Electronic Publication: 2024 Aug 15.
DOI: 10.1016/j.jviromet.2024.115008
Abstrakt: Infectious hematopoietic necrosis virus (IHNV) severely and lethally infects salmonid fish, including Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) worldwide. Rapid and accurate viral detection is crucial for preventing pathogen spread and minimizing damage. Although several IHNV detection assays have been developed, their analytical and diagnostic performances have not been evaluated and field usability assessments have not been completely validated. Here, we developed a reverse-transcription cross-priming amplification-based lateral flow assay (RT-CPA-LFA) and validated its diagnostic performance. To detect the IHNV, primers were designed based on the consensus sequence of the nucleocapsid (N) gene. Notably, when combined with a lateral flow dipstick, it could visualize the IHNV amplification products within 5 min and the detection limit of the developed RT-CPA-LFA was 3.28×10 5 copies/μL. The diagnostic sensitivity and specificity in fish samples (n=140) were 98.88 % and 96.08 %, respectively. Moreover, the IHNV detection rate by RT-CPA-LFA in dead rainbow trout artificially injected with the virus was 100 %, consistent with to the results obtained from second conventional and real-time PCR, indicating its applicability for rapid IHNV detection and presumptive IHN diagnosis during the endemic period.
Competing Interests: Declaration of Competing Interest The author declares no conflicts of interest
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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